Variants of the PapM polypeptide of bacteria of the Streptomyces genus
AssigneeAventis Pharma S.A.
InventorsNathalie Bamas-Jacques, Denis Thibaut, Alain Famechon
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Abstract
The present invention relates to novel variants of the PapM polypeptide of bacteria of the Streptomyces genus which possess a substrate selectivity and/or an improved effectiveness compared to the wild-type polypeptide. It also relates to the nucleic acids encoding these variants, to the microorganisms which incorporate these nucleic acids and to their use for producing B components of Streptogramins.
Claims10
A nucleic acid encoding a variant of the PapM polypeptide of bacteria of the Streptomyces genus comprising SEQ ID NO:2, selected from the group consisting of
- a polypeptide comprising the sequence of wild-type PapM polypeptide of S. pristinaespiralis modified by the replacement of residue Gly 249,
- a polypeptide comprising the sequence of wild-type PapM polypeptide of S. pristinaespiralis modified by the replacement of residue Thr 192,
- a polypeptide comprising the sequence of wild-type PapM polypeptide of S. pristinaespiralis modified by the replacement of residues Gly 249 and Thr 192, and
- wherein the variant of the PapM polypeptide
- catalyzes at least one of the methylation of PAPA to MMPAPA and the methylation of MMPAPA to DMPAPA and
- exhibits an efficiency of methylation, as defined by the ratio Kcat/Km, which differs from the efficiency of methylation of the wild-type PapM polypeptide of S. pristinaespiralis for at least one of the methylations of PAPA or MMPAPA.
A nucleic acid comprising SEQ ID NO. 1 modified by mutation for the replacement of at least one of Gly 249 or Thr 192 and optionally modified by the replacement of at least one codon with an alternate codon encoding the same amino acid due to the degeneracy of the genetic code.
Dependencies are inferred from the claim text.